Lentivirus-based Immunogene Therapy for HIV Infection

描述(由申请人提供)：该提案的中心主题是继续测试慢病毒工程T细胞在HIV感染中的安全性和有效性。这项提议延续了我们团队近年来在利用过继T细胞转移开发HIV免疫修复以及开发慢病毒载体来设计HIV内在耐药性方面取得的进展。我们最近完成了世界上第一个慢病毒基因转移的初步测试。前五个课题的结果令人鼓舞，值得进行正式研究，以优化本RFA下的方法。这项建议代表了学术界和私营部门之间强有力的和富有成效的互动，这一互动非常积极地将慢病毒载体技术，特别是T细胞基因转移疗法，带到了床边，并以安全和严格的方式进行。总体目标是通过测试基于实验室的假说，谨慎地建立安全性或我们的创新临床试验，同时为随后的艾滋病毒/艾滋病免疫基因转移试验进行额外的慢病毒工程基础研究。这项建议的内容如下：临床试验引擎：针对艾滋病毒的慢病毒工程T细胞(项目1，C.六月和P.Tebas)。该项目将在L/11期试验中测试表达反义艾滋病毒的第一代和第二代慢病毒载体的安全性和有效性以及贩运。慢病毒工程的CD4T细胞对宿主病毒关系的影响将使用独特的患者样本来确定。慢病毒载体整合在临床(项目2，F.Bushman)：将对慢病毒整合进行全面评估，以预测慢病毒工程T细胞的长期安全性。工程先天T细胞对艾滋病毒感染的抵抗力(项目3，J.Riley和R.Dom)：将通过使用锌指核酸酶靶向TrimSA和其他新靶点来开发改进的慢病毒载体，并将使用SCID-Hu模型来选择用于临床测试的优化载体。该计划由3个核心支持：核心A，慢病毒GLP和GMP载体核心；核心B，细胞工程核心；核心C，管理和生物统计学核心。私营部门与VIRxsys公司和Sangamo生物科学公司建立了互动关系。这些研究将共同提供针对HIIV/AIDS的慢病毒载体的安全性和抗病毒效果的全面评估，并将通过为这类新的和令人兴奋的载体提供关键的安全数据来推动该领域的发展。 项目1：临床试验引擎：针对HIV的慢病毒工程T细胞(6月，C.H.) 描述(由申请人提供)：过继转移的T淋巴细胞对HIV感染具有治疗前景的原则已经确立。我们的长期目标是确定输注慢病毒工程T细胞的安全性，并测试第二代转基因的安全性和改善的抗病毒效果。我们的长期目标是消除艾滋病毒感染患者每天服用抗病毒药物的需要。在最近完成的一项第一阶段初步研究中，我们证明了单次输注慢病毒工程的自体CD4T细胞用于晚期抗HAART HIV感染的HIV感染者的安全性和可行性。到目前为止，还没有插入离子突变的证据，其中一名受试者的病毒载量有所减少。转基因T细胞的植入性和持久性令人满意，表明VSV-G伪型HIV慢病毒载体系统是非免疫原性的。基于我们以前对共刺激的CD4T细胞的研究，我们现在假设，多次输注表达VRX496反义env基因的慢病毒工程自体CD4T细胞将导致持续和更高水平的植入。我们进一步假设转基因将产生抗病毒作用。两个临床试验被用来检验这些假说。首先，我们将在HAART抑制病毒复制的患者中进行多剂量L/11期研究。将进行有组织的治疗中断以评估抗病毒效果，并将使用淋巴活检来确定工程CD4T细胞的组织转运。在试验#2中，我们将测试项目3中开发的慢病毒载体，该载体表达更有效的抗病毒产品。我们将比较用第二代载体转导的T细胞和用试验1中测试的原始VRX496载体转导的T细胞的相对存活率。这些试验将共同代表慢病毒工程T细胞作为治疗HIV-1感染的有效抗病毒疗法的潜力的第一次正式疗效测试。该项目与项目2和项目3相互作用，该项目依赖于cGMP慢病毒载体制造和临床级T细胞扩增和转导技术的核心A和B。

DESCRIPTION (provided by applicant): The central theme of the proposal is to continue testing lentiviral engineered T cells for safety and efficacy in HIV infection. This proposal continues the progress made recent years by our group in developing immune restoration for HIV using adoptive T cell transfer and the development of lentiviral vectors to engineer intrinsic HIV resistance. We have recently completed the world's first pilot test of lentiviral gene transfer in humans. Results from the first five subjects are encouraging and warrant formal study to optimize the approach under this RFA. This proposal represents a strong and productive interaction between academia and the private sector that is highly motivated in bringing lentiviral vector technology, in general, and T cell gene transfer therapy in particular, to the bedside, and to do this in a safe and rigorous fashion. The overall goal is to carefully establish the safety or our innovative clinical trials by testing laboratory-based hypotheses, while pursuing additional basic investigation of lentiviral engineering for subsequent immunogene transfer trials for HIV/AIDS. The elements of this proposal are as follows:  Clinical Trials Engine: Lentiviral Engineered T Cells for HIV (Project 1, C. June and P. Tebas). This project will test first and second generation lentiviral vectors that express anti-sense HIV for safety and efficacy and trafficking in phase l/ll trials. Effects of lentiviral engineered CD4 T cells on the host virus relationship will be determined using unique patient samples.  Lentiviral Vector Integration in the Clinic (Project 2, F. Bushman): A comprehensive evaluation of lentiviral integration will be done to predict long term safety of lentiviral engineered T cells.  Engineering Innate T Cell Resistance to HIV Infection (Project 3, J. Riley and R. Doms): Improved lentiviral vectors will be developed by targeting TRIMSa and other novel targets using zinc finger nucleases, and a SCID-hu model will be used to select an optimized vector for clinical testing. The program is supported by 3 cores: Core A, the Lentiviral GLP and GMP Vector Core; Core B, the Cell Engineering Core, and Core C, the Administrative and Biostatistics Core. Private sector interactions are established with ViRxSYS Corp. and Sangamo BioSciences, Inc. Together these studies will provide a comprehensive evaluation of the safety and antiviral efficacy of lentiviral vectors for HIIV/AIDS, and they will move the field forward by providing critical safety data for this new and exciting class of vectors. PROJECT 1: Clinical trials engine: lentiviral engineered T cells for HIV (June, C. H.) DESCRIPTION (provided by applicant): The principle that adoptively transferred T lymphocytes have therapeutic promise for HIV infection is well established. Our long range goals are to establish the safety of infusions of lentiviral engineered T cells, and to test second generation transgenes for safety and improved antiviral efficacy. Our long range objective is to obviate the need to take daily antiviral medications in patients with HIV infection. In a recently completed phase I pilot study, we have demonstrated the safety and feasibility of a single infusion of lentiviral engineered autologous CD4 T cells when administered to HIV infected subjects with late-stage, HAART resistant HIV infection. To date, there is no evidence of insert ional mutagenesis, and one subject has experienced a reduction in viral load. The engraftment and persistence of the gene-modified T cells is satisfactory and suggests that the VSV-G pseudotyped HIV-based lentiviral vector system is nonimmunogenic. Based on our previous studies of costimulated CD4 T cells, we now hypothesize that multiple infusions of lentiviral engineered autologous CD4 T cells that express the VRX496 antisense env transgene will lead to a sustained and higher level engraftment. We further hypothesize that the transgene will confer antiviral effects. Two clinical trials are proposed to test these hypotheses. First, we will perform a multiple dose phase l/ll study in patients whose viral replication is suppressed on HAART. Structured treatment interruption will be carried out to assess antiviral efficacy, and lymphoid biopsies will be used to determine tissue trafficking of the engineered CD4 T cells. In trial #2 we will test a lentiviral vector developed in project 3 that expresses a more potent antiviral product. We will compare the relative survival of the T cells transduced with the second generation vector to cells transduced with the original VRX496 vector tested in trial #1. Together, these trials will represent the first formal efficacy tests of lentiviral engineered T cells for their potential to serve as a potent antiviral therapy for treatment of HIV-1 infection. This project interacts with projects 2, and 3, and the project relies on Cores A and B for cGMP lentiviral vector manufacturing and for clinical grade T cell expansion and transduction technology.